ThermoGen proposes to develop a library of stable esterases and lipases from a large bank of thermostable microorganisms. These new biocatalysts may be used in chiral synthesis of new and safer anticancer and antiviral pharmaceuticals. In Phase I we have identified a number of esterases and lipases with different substrate specificities from 76 different thermophilic bacterial strains. At least 35 esterase and 55 lipase activities were identified. Substrate specificity has been determined for a subset of the best characterized esterases. Two esterases have been purified and we have performed preliminary characterization with respect to kinetic properties, temperature optima, pH optima, and organic solvent tolerance. The N- terminal sequence of one esterase has been determined. Several of the esterases have been evaluated by chemists from pharmaceutical companies. In Phase II, ThermoGen will continue to expand the size of both the organism bank and the esterase and lipase library. Enzymes will be evaluated by a rapid screening system and key enzymes will be fully characterized and developed for commercial use. Prototype screening kits will be developed and commercialized. In Phase III commercial development will be performed by offering enzyme screening services, or, selling enzyme screening kits and/or bulk thermostable biocatalysts. PROPOSED COMMERCIAL APPLICATION: ThermoGen proposes to develop a large series of esterases and lipases as a library for chiral chemical synthesis applications. A subset of these biocatalysts with high stability and varied reaction characteristics will be produced for specific medical and industrial synthesis applications.